Turpentine Ointment for the Treatment of Folliculitis: An Open, Prospective, Randomized, Placebo- and Comparator-Controlled Multicenter Trial.

INTRODUCTION: Folliculitis is a painful infection and inflammation of the hair follicles, mostly caused by bacterial, fungal, or, more rarely, viral infections. Turpentine derivatives have been used traditionally to treat various skin infections and could thus also be effective in treating folliculitis. We carried out an open, prospective, randomized, placebo- and comparator-controlled multicenter trial to evaluate the efficacy and safety of an ointment containing pine turpentine oil, larch turpentine, and eucalyptus oil in the treatment of acute folliculitis. METHODS: Seventy outpatients with acute folliculitis were treated with the turpentine ointment, a comparator (povidone iodine solution), or a placebo (Vaseline) for 7 days. Photographs of the affected skin areas were taken by the physicians at four visits and by the patients on a daily basis. Photographs were evaluated by blinded observers. Primary efficacy endpoint was the change in total hair follicle lesion counts. Secondary endpoints included the evolution of the lesion counts in the course of the study, responder rate (improvement of follicle lesions by at least one count), and the patient's global assessment. Safety endpoints were the tolerability of the treatments and adverse event recording. RESULTS: A decrease of follicle lesions counts was detected for both active treatments but not for placebo, but the differences among groups were not statistically significant. As for the secondary endpoints, the ointment showed statistically significant superiority over placebo for the evolution of the lesions during the course of the study (p = 0.017), the responder rate (p = 0.032), and the subjective efficacy assessment by patients (p = 0.029). All treatments were equally well tolerated, with a similar number of treatment-emergent adverse events. CONCLUSION: The turpentine ointment is an effective and safe option for the treatment of folliculitis.

Turpentine Ointment in Bacterial Skin Infections: A Randomized, Placebo-Controlled, Double-Blind Clinical Trial.

BACKGROUND: Turpentine-containing substances are considered effective in treating cutaneous bacterial infections, but reliable clinical data are scant. OBJECTIVE: We investigated the efficacy and safety of an ointment containing larch turpentine (from Larix decidua), eucalyptus oil (from Eucalyptus globulus), and turpentine oil (from Pinus pinaster) in outpatients with painful skin abscesses in a randomized, placebo-controlled, double-blind study. INTERVENTION: 116 outpatients with skin abscesses used verum or placebo for 10 days. Sum score of the patient's discomforts, changes in abscess size, rate of therapeutic success, and complete healing served as outcome parameters. RESULTS: Fifty-four patients were treated with verum and 56 with placebo. According to the patient's discomfort sum score, patients in the verum group showed a better improvement compared to the placebo group (7.3 vs. 4.7; p = 0.024), and subjective assessment by the investigators revealed a higher treatment success rate after verum (70% vs. 48%; p = 0.021). Complete healing was documented in 67% of the patients receiving verum versus 46% in the placebo group (p = 0.037). There was a positive trend toward a larger decrease in the abscess sizes in the verum group compared to the placebo group (p = 0.07). CONCLUSION: The ointment studied is an effective and safe option for the treatment of bacterial skin diseases.

[Homöopathisch-phytotherapeutische Behandlung des Reizdarmsyndroms mit Magen-Darm-Entoxin N®: Eine Anwendungsbeobachtung].

Hintergrund: In dieser Anwendungsbeobachtung wird zum ersten Mal die Behandlung des Reizdarmsyndroms (RDS) mit einem homöopathisch-phytotherapeutischen Komplexpräparat beschrieben. Methodik: Ziel der 6-wöchigen Therapie mit dem Magen-Darm-Entoxin N® war die Reduzierung der RDS-Symptomatik sowie die Verbesserung der Lebensqualität. Zielkriterien waren die Veränderungen in der Irritable Bowel Syndrome - Severity Scoring System (IBS-SSS) und der Irritable Bowel Syndrome - Quality-of-Life Scale (IBS-QoL). Die Patient/innen (N = 41; Alter 44,0 ± 15,74 Jahre) wurden zu gleichen Teilen in einer Hausarztpraxis (N = 20) und einer Heilpraktikerpraxis (N = 21) rekrutiert. Ergebnisse: Der IBS-QoL-Score verringerte sich signifikant (prä: 35,9 ± 16,3; post: 20,1 ± 13,4; t = 8,504; p < 0,001). Die Effektstärke betrug 1,34 (Cohens d). Der IBS-SSS-Score verringerte sich ebenfalls signifikant (prä: 239,4 ± 83,4; post: 123,7 ± 80,9; t = 7,825; p < 0,001) mit einer Effektstärke von d = 1,24. Die Neben- und Wechselwirkungen waren minimal und signifikante Unterschiede zwischen beiden Praxen wurden nicht gefunden. Schlussfolgerungen: Magen-Darm-Entoxin N® ist eine sichere und sinnvolle Therapieoption bei der Behandlung des RDS. Allerdings sollten randomisierte kontrollierte Studien folgen, um die Spezifizität der Ergebnisse dieser Anwendungsbeobachtung zu stützen.

Characterization of the gene cluster CYP264B1-geoA from Sorangium cellulosum So ce56: biosynthesis of (+)-eremophilene and its hydroxylation.

Terpenoids can be found in almost all forms of life; however, the biosynthesis of bacterial terpenoids has not been intensively studied. This study reports the identification and functional characterization of the gene cluster CYP264B1-geoA from Sorangium cellulosum So ce56. Expression of the enzymes and synthesis of their products for NMR analysis and X-ray diffraction were carried out by employing an Escherichia coli whole-cell conversion system that provides the geoA substrate farnesyl pyrophosphate through simultaneous overexpression of the mevalonate pathway genes. The geoA product was identified as a novel sesquiterpene, and assigned NMR signals unambiguously proved that geoA is an (+)-eremophilene synthase. The very tight binding of (+)-eremophilene (∼0.40 µM), which is also available in S. cellulosum So ce56, and its oxidation by CYP264B1 suggest that the CYP264B1-geoA gene cluster is required for the biosynthesis of (+)-eremophilene derivatives.

A diterpene synthase from the clary sage Salvia sclarea catalyzes the cyclization of geranylgeranyl diphosphate to (8R)-hydroxy-copalyl diphosphate.

The bicyclic diterpene (-)-sclareol is accumulated in glandular trichomes in Salvia sclarea (Schmiderer et al., 2008), and is a major terpenoid component of this plant species. It is used as the starting material for Ambrox synthesis, a synthetic ambergris analog used in the flavor and fragrance industry. In order to investigate the formation of sclareol, cDNA prepared from secretory cells of glandular trichomes from S. sclarea inflorescence were randomly sequenced. A putative copalyl diphosphate synthase encoding EST, SscTPS1, was functionally expressed in Escherichia coli. Whereas reaction of geranylgeranyl diphosphate with the putative copalyl diphosphate synthase followed by hydrolysis with alkaline phosphatase yielded a diastereomeric mixture of (13R)- and (13S)-manoyl oxide, HCl hydrolysis yielded (-)-sclareol (1) and 13-epi-sclareol as products. The product of the reaction of SscTPS1 with geranylgeranyl diphosphate was subjected to analysis by LC-negative ion ESI-MS/MS without prior hydrolysis. EPI scans were consistent with copalyl diphosphate to which 18 mass units had added (m/z 467 [M+H](-)). The enzymatic reaction was also carried out in the presence of 60% H2(18)O. LC-negative ion ESI-MS/MS analysis established an additional reaction product consistent with the incorporation of (18)O. Incubation in the presence of 60% (2)H2O resulted in the incorporation of one deuterium atom. These results suggest water capture of the carbocation intermediate, which is known to occur in reactions catalyzed by monoterpene synthases, but has been described only several times for diterpene synthases.

Changing the regioselectivity of a P450 from C15 to C11 hydroxylation of progesterone.

CYP106A2 is known as a 15ß-hydroxylase, but also shows minor 11α-hydroxylase activity for progesterone. 11α-Hydroxyprogesterone is an important pharmaceutical compound with anti-androgenic and blood-pressure-regulating activity. This work therefore focused on directing the regioselectivity of the enzyme towards hydroxylation at position 11 in the C ring of the steroid through a combination of saturation mutagenesis and rational site-directed mutagenesis. With the aid of data from a homology model of CYP106A2 containing docked progesterone, together with site-directed mutagenesis of active-site residues (Lisurek et al. ChemBioChem 2008, 9, 1439-1449), a saturation mutagenesis library at positions A395 and G397 was created. Screening of the library identified the mutants A395I and A395W/G397K as having 11α-hydroxylase activities 8.9 and 11.5 times higher than that of the wild type (WT). In the next step, additional mutations were integrated by a rational site-directed mutagenesis approach to increase the catalytic efficiency. Of the 40 candidates analyzed, the mutants A106T/A395I, A106T/A395I/R409L, and T89N/A395I turned out to display increased 11α-hydroxylase selectivities and activities relative to the WT (14.3-, 12.6-, and 11.8-fold increases in selectivity and 39.3-, 108-, and 24.4- in k(cat)/K(m)). In the last step of the study, the best mutants were applied in a whole-cell biotransformation. In these experiments the production (percentage) of 15ß-hydroxyprogesterone decreased from 50.4 % (wild type) to 4.8 % (mutant T89N/A395I), whereas that of 11α-hydroxyprogesterone increased from 27.7 to 80.9 %, thus demonstrating an impressive regioselectivity.

Molecular and structural basis of metabolic diversity mediated by prenyldiphosphate converting enzymes.

General thermodynamic calculations using the semiempiric PM3 method have led to the conclusion that prenyldiphosphate converting enzymes require at least one divalent metal cation for the activation and cleavage of the diphosphate-prenyl ester bond, or they must provide structural elements for the efficient stabilization of the intermediate prenyl cation. The most important common structural features, which guide the product specificity in both terpene synthases and aromatic prenyl transferases are aromatic amino acid side chains, which stabilize prenyl cations by cation-pi interactions. In the case of aromatic prenyl transferases, a proton abstraction from the phenolic hydroxyl group of the second substrate will enhance the electron density in the phenolic ortho-position at which initial prenylation of the aromatic compound usually occurs. A model of the structure of the integral transmembrane-bound aromatic prenyl transferase UbiA was developed, which currently represents the first structural insight into this group of prenylating enzymes with a fold different from most other aromatic prenyl transferases. Based on this model, the structure-activity relationships and mechanistic aspects of related proteins, for example those of Lithospermum erythrorhizon or the enzyme AuaA from Stigmatella aurantiaca involved in the aurachin biosynthesis, were elucidated. The high similarity of this group of aromatic prenyltransferases to 5-epi-aristolochene synthase is an indication of an evolutionary relationship with terpene synthases (cyclases). This is further supported by the conserved DxxxD motif found in both protein families. In contrast, there is no such relationship to the aromatic prenyl transferases with an ABBA-fold, such as NphB, or to any other known family of prenyl converting enzymes. Therefore, it is possible that these two groups might have different evolutionary ancestors.